1 reply to this topic

[Sharon Pek]

Hi,

I'm a beginner in mitochondria extraction and I'm hoping to get some advice.
I've got the kit from pierce, which using a reagent based method for extraction (no homogeniser needed)

Normally, what's a good amount of starting material from cell lines?

I've started with a T75, fully confluent, but I didnt seem to get any mitochondria pellet?

Thanks

[Carlton H]

Sharon Pek, on Apr 24 2009, 03:46 AM, said:

Normally, what's a good amount of starting material from cell lines?

Depends on how much mitochondia you need.

If you're using very few cells, then perhaps the pellet is too small to see.  Have a good light microscope?  If so, take a look.

I hate to suggest this, because I know that people buy the kits specifically to avoid doing this, but if you've tried the kit a few times (scaling up the amount of cells used) and haven't got anything perhaps you should double-check using the homogenization method.  If you use two methods and you still can't get mitochondria, then it's probably not a problem with the method and you can start troubleshooting other aspects of your experiment.

Let me know how things work out.

Cheers,
-Carlton